Shaping the Microbial Landscape: Parasitoid-Driven Modifications of Bactrocera dorsalis Microbiota.

Koinobiont endoparasitoids regulate the physiology of their hosts through altering host immuno-metabolic responses, processes which function in tandem to shape the composition of the microbiota of these hosts. Here, we employed 16S rRNA and ITS amplicon sequencing to investigate whether parasitization by the parasitoid wasps, Diachasmimorpha longicaudata (Ashmaed) (Hymenoptera: Braconidae) and Psyttalia cosyrae (Wilkinson) (Hymenoptera: Braconidae), induces gut dysbiosis and differentially alter the gut microbial (bacteria and fungi) communities of an important horticultural pest, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). We further investigated the composition of bacterial communities of adult D. longicaudata and P. cosyrae to ascertain whether the adult parasitoids and parasitized host larvae share microbial taxa through transmission. We demonstrated that parasitism by D. longicaudata induced significant gut perturbations, resulting in the colonization and increased relative abundance of pathogenic gut bacteria. Some pathogenic bacteria like Stenotrophomonas and Morganella were detected in both the guts of D. longicaudata-parasitized B. dorsalis larvae and adult D. longicaudata wasps, suggesting a horizontal transfer of microbes from the parasitoid to the host. The bacterial community of P. cosyrae adult wasps was dominated by Arsenophonus nasoniae, whereas that of D. longicaudata adults was dominated by Paucibater spp. and Pseudomonas spp. Parasitization by either parasitoid wasp was associated with an overall reduction in fungal diversity and evenness. These findings indicate that unlike P. cosyrae which is avirulent to B. dorsalis, parasitization by D. longicaudata induces shifts in the gut bacteriome of B. dorsalis larvae to a pathobiont-dominated community. This mechanism possibly enhances its virulence against the pest, further supporting its candidacy as an effective biocontrol agent of this frugivorous tephritid fruit fly pest.

Friend or Foe: Symbiotic Bacteria in Bactrocera dorsalis-Parasitoid Associations.

Parasitoids are promising biocontrol agents of the devastating fruit fly, Bactrocera dorsalis. However, parasitoid performance is a function of several factors, including host-associated symbiotic bacteria. Providencia alcalifaciens, Citrobacter freundii, and Lactococcus lactis are among the symbiotic bacteria commonly associated with B. dorsalis, and they influence the eco-physiological functioning of this pest. However, whether these bacteria influence the interaction between this pest and its parasitoids is unknown. This study sought to elucidate the nature of the interaction of the parasitoids, Fopius arisanus, Diachasmimorpha longicaudata, and Psyttlia cosyrae with B. dorsalis as mediated by symbiotic bacteria. Three types of fly lines were used: axenic, symbiotic, and bacteria-mono-associated (Lactococcus lactis, Providencia alcalifaciens, and Citrobacter freundii). The suitable stages of each fly line were exposed to the respective parasitoid species and reared until the emergence of adult flies/parasitoids. Thereafter, data on the emergence and parasitoid fitness traits were recorded. No wasps emerged from the fly lines exposed to P. cosyrae. The highest emergence of F. arisanus and D. longicaudata was recorded in the L. lactis fly lines. The parasitoid progeny from the L. lactis and P. alcalifaciens fly lines had the longest developmental time and the largest body size. Conversely, parasitoid fecundity was significantly lower in the L. lactis lines, whereas the P. alcalifaciens lines significantly improved fecundity. These results elucidate some effects of bacterial symbionts on host-parasitoid interactions and their potential in enhancing parasitoid-oriented management strategies against B. dorsalis.

Screening the immunotoxicity of different food preservative agents on the model organism Galleria mellonella L. (Lepidoptera: Pyralidae) larvae.

Immunotoxic effects of sodium benzoate (SB, E211), sodium nitrate (SNa, E251), and sodium nitrite (SNi, E250), a few of the most common food preservatives, on the model organism Galleria mellonella L. (Lepidoptera: Pyralidae) larvae were investigated in this study. The last instar larvae were used for all experimental analyses. For this purpose, median lethal doses of SB, SNa, and SNi were applied to the larvae by the force-feeding method. We found that force-feeding G. mellonella larvae with SB, SNa, and SNi significantly reduced the larval total hemocyte counts, prohemocyte, and granulocyte ratios but increased plasmatocyte, spherulocyte, and oenocyte ratios, as well as the hemocyte mitotic indices and micronucleus frequency. The spreading ability of hemocytes and hemocyte-mediated immune responses were lower in the SB, SNa-, and SNi-treated larval groups compared to controls. Apoptotic indices were higher in all larval groups treated with food preservatives, but increments in necrotic indices were only significantly higher in SNi-treated larvae compared to controls. Our research shows that SB, SNa, and SNi have immunotoxic and cytotoxic potential on G. mellonella larvae. Thus, we suggest that G. mellonella larvae can be used as preliminary in vivo models to screen the immunotoxic effects of food preservative agents.

Differential immune responses in new and old fruit fly-parasitoid associations: Implications for their management.

The oriental fruit fly, Bactrocera dorsalis (Hendel), and marula fruit fly, Ceratitis cosyra (Walker), are major fruit-infesting tephritids across sub-Saharan Africa. Biological control of these pests using parasitic wasps has been widely adopted but with varying levels of success. Most studies investigating host-parasitoid models have focused on functional and evolutionary aspects leaving a knowledge gap about the physiological mechanisms underpinning the efficacy of parasitoids as biocontrol agents of tephritids. To better understand these physiological mechanisms, we investigated changes in the cellular immune responses of C. cosyra and B. dorsalis when exposed to the parasitic wasps, Diachasmimorpha longicaudata (Ashmaed) and Psyttalia cosyrae (Wilkinson). We found that B. dorsalis was more resistant to parasitisation, had a higher hemocyte count, and encapsulated more parasitoid eggs compared to C. cosyra, achieving up to 100% encapsulation when exposed to P. cosyrae. Exposing B. dorsalis to either parasitoid species induced the formation of a rare cell type, the giant multinucleated hemocyte, which was not observed in C. cosyra. Furthermore, compared to P. cosyrae-parasitized larvae, those of both host species parasitized by D. longicaudata had lower encapsulation rates, hemocyte counts and spreading abilities and yielded a higher number of parasitoid progeny with the highest parasitoid emergence (72.13%) recorded in C. cosyra. These results demonstrate that cellular immune responses are central to host-parasitoid interaction in tephritid fruit flies and further suggest that D. longicaudata presents greater potential as a biocontrol agent of B. dorsalis and C. cosyra in horticultural cropping systems.

Immunotoxic effects of force-fed ethephon on model organism Galleria mellonella (Lepidoptera: Pyralidae).

Incorporation of chemical substances like plant growth regulators in agricultural practices to boost production has become inevitable; thus, they have accumulated in the environment in tremendous amounts. However, due to their nonselective nature, they affect several components of the ecosystem like the invertebrates. In this study, therefore, the effects of force-fed Ethephon on the cellular mediated immune system of model insect G. mellonella larvae were investigated using the lethal doses LD25 and LD50 determined in a previous study. Our results indicated that treating G. mellonella larvae with ETF significantly reduces the number of circulating hemocytes and also reduces the number of live cells while increasing the apoptotic and necrotic cell ratios at all doses. Additionally, ETF increased the number of spherulocytes, oenocytes and prohemocytes as well as the mitotic indices while reducing the number of granulocytes in circulation but did not alter the number of plasmatocytes. Moreover, the in vivo encapsulation assays showed significant suppression of the encapsulation abilities of the ETF treated G. mellonella larval hemocytes at both ETF doses. The findings of the current study are indicative of the ecotoxic effects that may arise due to ETF and that its usage should be controlled or monitored as it poses major threats to several organisms and the ecosystem at large.

Boric acid-induced immunotoxicity and genotoxicity in model insect Galleria mellonella L. (Lepidoptera: Pyralidae).

Boric acid (BA) is widely used in various industrial process and can be accessed to nontarget organisms. This study aimed to investigate the insecticidal effects of BA and its toxic activities with respect to immunologic and genotoxic effects using Galleria mellonella larvae as a model. BA concentrations (78.125-10,000 ppm) were administrated to the larvae using the feeding method. Concentration-dependent mortality was observed in all larval groups. Probit analysis revealed LC30 , LC50 , and LC70 values to be 112.4, 320.1, and 911.4 ppm, respectively. These concentrations were used in all bioassays. Drastic reductions in total hemocyte counts along with changes in differential hemocyte counts were observed following BA treatment. Cell viability assays showed dose-dependent reductions in viable cells and an increase in the necrotic and apoptotic ratios after BA treatment. However, mitotic indices of larval hemocytes did not change at all BA concentrations. The cytotoxic effect of BA led to a significant reduction in cellular immune responses such as encapsulation, melanization, and nodulation activities of treated larvae. While BA increased micronucleus ratios at the highest concentration, comet parameters indicating DNA damage increased in G. mellonella larval hemocytes at all concentrations. These report that BA suppresses the immune system of G. mellonella and also poses risks of genotoxicity at high concentrations.